Fluorogenic assay for penicillin G acylase activity.

M Ninkovic; D Riester; F Wirsching; R Dietrich; A Schwienhorst

doi:10.1006/abio.2001.5078

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Fluorogenic assay for penicillin G acylase activity.

M Ninkovic·D Riester·F Wirsching·R Dietrich·A Schwienhorst

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Abstract

A simple, highly sensitive, and rapid assay for high-throughput screening of penicillin G acylase-producing bacteria is presented. The method is based on the specific release of fluorescent 7-amino-4-methyl-coumarin through cleavage of phenylacetyl-4-methyl-coumaryl-7-amide by penicillin G acylase. The present method is suitable for screening pure enzymes as well as various penicillin G acylases like those from Escherichia coli, Proteus rettgeri, and Kluyvera citrophila in cell extracts. In addition, the new substrate was used for rapid assay of amidase activity in nondenaturing polyacrylamide gels.

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Coumarins Electrophoresis, Polyacrylamide Gel Escherichia coli Fluorescent Dyes Indicators and Reagents Penicillin Amidase Recombinant Fusion Proteins Sensitivity and Specificity Spectrometry, Fluorescence Time Factors Journal Article Research Support, Non-U.S. Gov't

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M Ninkovic, D Riester, F Wirsching, R Dietrich, A Schwienhorst. 2001-05-15. Fluorogenic assay for penicillin G acylase activity.. https://doi.org/10.1006/abio.2001.5078

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